胎猪胰腺干细胞的分离与鉴定  

Isolation and Identification of Porcine Fetus Pancreatic Stem Cells

作　　者：王赟[1] 赵婷[1] 乔海[1] 杨春荣[1] 张翊华[1] 王华岩[1] 窦忠英[1] 

机构地区：[1]西北农林科技大学国家干细胞生物技术中心陕西分中心,杨凌712100

出　　处：《农业生物技术学报》2007年第4期622-627,共6页Journal of Agricultural Biotechnology

基　　金：国家高技术研究与发展计划(863)项目(No.2005AA219050);教育部重大项目(No.03160);国家自然科学基金(No.30671067);陕西省重大科技专项(No.2006kz05-G1)资助

摘　　要：从胎猪胰腺分离得到1株细胞,该细胞经免疫组化检测表达胰肠同源域因子1(pancreatic duodenal homeobox-1,PDX-1)、葡萄糖转运子2(glucose transporter-2,GLUT-2)、增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)、胰高血糖素(glucagon)、生长抑素(somatostatin)、胰多肽(pancreatic polypeptide)和弱表达巢蛋白(nestin),目前细胞已传至58代。其在体外生长类似神经样细胞生长形态,经诱导后可分化为具有三维立体结构的类胰岛细胞团(islet-like cell clusters,ICCs)。成团后细胞强表达胰岛素(insulin)、角蛋白19(cytokeratin 19,CK19)等标志物,放免测定显示诱导成团后的细胞胰岛素分泌量增加,最高达30.3±7.5μIU/mL,显著高于未经诱导的细胞(6.3±1.2μIU/mL,P<0.05)。经双硫腙(DTZ)染色呈亮红色,这些结果说明胰腺内存在着多潜能干细胞。实验分离到胎猪胰腺干细胞,经诱导分化可形成有功能的胰岛样细胞团并释放胰岛素,有望为进一步研究异种胰腺干细胞移植治疗糖尿病提供材料。Cells derived from porcine fetus were isolated and sub-cloned over 58 passages during 13 months and identified as pancreatic stem cells by immunohistochemistry, which were morphologicaly similar with neuron-like cells and were strongly positive for pancreatic duodenal homeobox-1 （PDX-1）,glucose transporter-2 （GLUT-2）,proliferating cell nuclear antigen （PCNA）, glucagon, somatostatin and pancreatic polypeptide, respectively, but weakly positive for nestin. After in vitro induction these cells differentiated into islet-like cell clusters（ICCs） with three-dimension structure,and some markers - insulin and cytokeratin 19 （CK 19） were strongly expressed, in ICCs. By means of radioimmunoassay（RIA）, ICCs had an increase in secretion of insulin ,whose highest level was 30.3± 7.5 μIU/mL, significantly higher than that of control group （6.3± 1.2 μIU/mL）（P 〈0.05）. Furthermore, ICCs were brightened for dithizone （DTZ） staining.These results indicated that there are multipotent stem cells in pancreas. The present study offers an approach to isolate and expand pancreatic stem cells from porcine fetus, and differentiate them into functional ICCs in vitro. It likely becomes a new source for diabetes therapy through xenotransplantation of pancreatic stem cells.

关 键 词：胎猪 胰岛 胰腺干细胞 糖尿病 

分 类 号：S185[农业科学—农业基础科学]